Development of a Reproducible Model of Parenteral Nutrition-Associated Liver Disease in Rats

Abstract

Purpose For patients with intestinal failure, parenteral nutrition (PN) is a life-saving therapy. Unfortunately, hepatic dysfunction will occur in 40 to 60% of children on long-term PN. While the hepatic dysfunction is likely multifactorial, one important chemical component of the disease may be aluminum contamination of the PN. Previous studies have shown a reduction in liver injury by decreasing the aluminum concentration in PN in a pig model. We sought to develop a rat model of PN-associated liver disease (PNALD) with parenteral aluminum.

Methods Adult Sprague–Dawley rats had intravenous long-term catheters placed. The control group underwent daily injections of saline. The study rats had daily injections of either 2 or 3 mg/kg aluminum chloride (AlCl₃). At the end of 4 weeks, the rats were euthanized and liver and blood samples were taken. The livers were analyzed and graded by a pathologist for histological evidence of liver degeneration and acute and chronic inflammation. The serum was analyzed for total bilirubin, alkaline phosphatase, alanine aminotransferase (ALT), and aspartate aminotransferase (AST).

Results There was no difference in serum values of total bilirubin, alkaline phosphatase, ALT, or AST. There was no difference in acute inflammation among the groups (1 [control], 1.2 [2 mg/kg], 1.1 [3 mg/kg]). The rats treated with parenteral aluminum had significantly more Kupffer cells than the control group (0.1 [control], 3 [2 mg/kg], 2.2 [3 mg/kg], p < 0.0001 [control vs. 2 mg/kg] and 0.0032 [control vs. 3 mg/kg]). There was also more liver degeneration in the parenteral aluminum groups than the control group (1 [control], 2 [2 mg/kg], 2.5 [3 mg/kg], p = 0.0341 [control vs. 2 mg/kg] and 0.009 [control vs. 3 mg/kg]). However, there was no difference between 2 and 3 mg/kg AlCl₃ for either variable.

Conclusion This study suggests that 4 weeks of parenteral aluminum can induce chronic inflammation and degeneration of the liver in rats. Therefore, we believe that daily injections of parenteral aluminum can produce a viable model of PNALD in rats. However, further studies are warranted, including measurement of serum aluminum levels in infants on PN.

Keywords

parenteral nutrition - cholestatic liver disease - rat

• References

• 1 Zambrano E, El-Hennawy M, Ehrenkranz RA, Zelterman D, Reyes-Múgica M. Total parenteral nutrition induced liver pathology: an autopsy series of 24 newborn cases. Pediatr Dev Pathol 2004; 7 (5) 425-432
• 2 Drongowski RA, Coran AG. An analysis of factors contributing to the development of total parenteral nutrition-induced cholestasis. JPEN J Parenter Enteral Nutr 1989; 13 (6) 586-589
• 3 Kubota A, Okada A, Nezu R, Kamata S, Imura K, Takagi Y. Hyperbilirubinemia in neonates associated with total parenteral nutrition. JPEN J Parenter Enteral Nutr 1988; 12 (6) 602-606
• 4 Ginn-Pease ME, Pantalos D, King DR. TPN-associated hyperbilirubinemia: a common problem in newborn surgical patients. J Pediatr Surg 1985; 20 (4) 436-439
• 5 Alemmari A, Miller GG, Arnold CJ, Zello GA. Parenteral aluminum induces liver injury in a newborn piglet model. J Pediatr Surg 2011; 46 (5) 883-887
• 6 Klein GL, Heyman MB, Lee TC , et al. Aluminum-associated hepatobiliary dysfunction in rats: relationships to dosage and duration of exposure. Pediatr Res 1988; 23 (3) 275-278
• 7 Demircan M, Ergün O, Coker C, Yilmaz F, Avanoglu S, Ozok G. Aluminum in total parenteral nutrition solutions produces portal inflammation in rats. J Pediatr Gastroenterol Nutr 1998; 26 (3) 274-278
• 8 Alemmari A, Miller GG, Bertolo RF , et al. Reduced aluminum contamination decreases parenteral nutrition associated liver injury. J Pediatr Surg 2012; 47 (5) 889-894
• 9 Li Z, Yang S, Lin H , et al. Probiotics and antibodies to TNF inhibit inflammatory activity and improve nonalcoholic fatty liver disease. Hepatology 2003; 37 (2) 343-350
• 10 Thatch KA, Katz MS, Haber MM, Schwartz MZ. Growth factor modulation of hepatic inflammation: a novel approach to the management of total parenteral nutrition-associated liver disease. J Pediatr Surg 2010; 45 (1) 89-94
• 11 ASCN/A.S.P.E.N. Working Group on Standards for Aluminum Content of Parenteral Nutrition Solutions. Parenteral drug products containing aluminum as an ingredient or a contaminant: response to Food and Drug Administration notice of intent and request for information. JPEN J Parenter Enteral Nutr 1991; 15 (2) 194-198
• 12 Moreno A, Domínguez C, Ballabriga A. Aluminum in the neonate related to parenteral nutrition. Acta Paediatr 1994; 83 (1) 25-29
• 13 Arnold CJ, Miller GG, Zello GA. Parenteral nutrition-associated cholestasis in neonates: the role of aluminum. Nutr Rev 2003; 61 (9) 306-310
• 14 Popińska K, Kierkuś J, Lyszkowska M , et al. Aluminum contamination of parenteral nutrition additives, amino acid solutions, and lipid emulsions. Nutrition 1999; 15 (9) 683-686
• 15 Vilahur G, Padro T, Badimon L. Atherosclerosis and thrombosis: insights from large animal models. J Biomed Biotechnol 2011; 2011: 907575
• 16 Thatch KA, Schwartz MZ, Yoo EY, Mendelson KG, Duke DS. Modulation of the inflammatory response and apoptosis using epidermal growth factor and hepatocyte growth factor in a liver injury model: a potential approach to the management and treatment of cholestatic liver disease. J Pediatr Surg 2008; 43 (12) 2169-2173
• 17 Yoshikawa A, Kaido T, Seto S , et al. Hepatocyte growth factor promotes liver regeneration with prompt improvement of hyperbilirubinemia in hepatectomized cholestatic rats. J Surg Res 1998; 78 (1) 54-59